HAPIscreen, a method for high-throughput aptamer identification.

Area of Science:

• Biotechnology
• Molecular Biology
• Genomics

Background:

• Aptamers are oligonucleotides with specific binding properties selected via SELEX (Systematic Evolution of Ligands by EXponential enrichment).
• Traditional SELEX involves time-consuming sequencing and individual binding evaluation, potentially missing high-affinity aptamers.
• A need exists for methods that overcome these limitations in aptamer discovery.

Purpose of the Study:

• To develop and validate a novel, high-throughput method for aptamer identification.
• To enable functional identification of high-affinity aptamers directly from SELEX pools.
• To reduce bias and time associated with traditional aptamer selection and characterization.

Main Methods:

• A homogeneous solution-based screening method using AlphaScreen technology was developed.
• The HAPIscreen (High throughput APtamer Identification screen) methodology analyzes binding properties without prior sequencing.
• The method was validated using known aptamers and applied to identify new aptamers against pre-microRNAs.

Main Results:

• HAPIscreen enables the functional identification of high-affinity aptamers.
• The methodology was successfully validated using RNA hairpin aptamers.
• New RNA aptamers targeting pre-microRNAs were identified from SELEX pools.

Conclusions:

• HAPIscreen is a faster and less biased alternative to traditional aptamer identification methods.
• This AlphaScreen-based methodology allows for screening larger numbers of aptamer candidates.
• HAPIscreen is adaptable for various targets and amenable to automation.